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Analysis of lysozyme in cheese by immunocapture mass spectrometry

机译:免疫捕获质谱法分析奶酪中的溶菌酶

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The enzyme lysozyme is used as a preservative to prevent late blowing of ripened cheese, caused by Clostridium tyrobutyricum. Since the enzyme is extracted from hen egg white, lysozyme has to be declared on food product labels as a potential allergen. Here, a method is reported that combines immunocapture purification and matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDITOF- MS) analysis for the detection of lysozyme in cheese samples. Cheese extracts were treated with magnetic particles coated with a monoclonal antibody directed against lysozyme. After immunocapture purification, lysozyme was detected by MALDI-TOF-MS. The limit of detection of the assay was about 5 mg/kg lysozyme in cheese. The method reliably distinguished between cheese samples which had been produced with and without lysozyme. Thus, the novel assay allows the reliable, sensitive, and specific detection of lysozyme in a food matrix. The assay could be easily adapted to other target peptides and proteins in complex food matrices and, therefore, has a broad application potential, e.g. for the analysis of allergens.
机译:溶菌酶被用作防腐剂,以防止由酪丁酸梭状芽胞杆菌引起的成熟奶酪的后期起泡。由于该酶是从鸡蛋清中提取的,因此必须在食品标签上声明溶菌酶是潜在的过敏原。在此,报告了一种结合了免疫捕获纯化和基质辅助激光解吸/电离飞行时间质谱(MALDITOF-MS)分析的方法来检测奶酪样品中的溶菌酶。奶酪提取物用涂有针对溶菌酶的单克隆抗体包被的磁性颗粒处理。免疫捕获纯化后,通过MALDI-TOF-MS检测溶菌酶。该测定的检测极限是奶酪中约5mg / kg的溶菌酶。该方法可靠地区分了有和没有溶菌酶的干酪样品。因此,该新颖的测定方法允许可靠,灵敏且特异性地检测食物基质中的溶菌酶。该测定法可以很容易地适应复杂食品基质中的其他靶标肽和蛋白质,因此具有广泛的应用潜力,例如。用于分析过敏原。

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